Biomarkers for Malignant Potential in Vocal Fold Leukoplakia: A State of the Art Review.

OBJECTIVE: To explore biomarkers that are candidates for understanding potential degeneration to malignancy of vocal fold leukoplakia (VFL), with the goal of guiding future diagnostic and treatment recommendations. DATA SOURCES: PubMed and Medline search engines. REVIEW METHODS: A systematic review was conducted by searching the following key words: vocal fold or laryngeal, coupled with leukoplakia or dysplasia, and combined with the term prognostic markers. We collated the biomarkers and their significance, followed by observing the power of their evidence by assessing the quality of the studies according to guidelines of tumor marker prognostic studies (REMARK). CONCLUSIONS: Prognostic biomarkers in the 16 studies are generally divided into 3 categories according to their biological roles: proliferation (Ki-67, CK-1 RS14024 SNP), cell cycle control (P53, p16, cyclin D1, p57kip2, interleukin-10 [IL-10], miR-10a, and miR-34c), cell adhesion, and invasion (neutrophil-to-lymphocyte ratio, OPN/CD44v6 axis, MMP-1, vascular endothelial growth factor A, MMP-9, serpin peptidase inhibitor 1, plasminogen activator, CTNN/B1, ß-catenin, NANOG, HERG1). The prognostic use of these biomarkers is limited due to the variable methodologies, study design, assay methods, and statistical analysis performed. IMPLICATIONS FOR PRACTICE: Prognostic factors in vocal fold leukoplakia have important clinical implications regarding the potential for malignant degeneration. Although further study is needed, the currently available evidence suggests that p53, p16, cyclin D1, IL-10, NLR, OPN and CD44v6, CTNNB1, and CTTN and FAK might be of particular interest in determining prognosis of VFL as related to malignancy. Future, large, well-designed, prospective studies are expected to determine the prognostic power of these biomarkers before their implementation in routine clinical practice.

An insight into crystallization test: A neoteric approach for screening premalignant and malignant lesions.

INTRODUCTION: Crystallization test is based on the principle that, when a salt crystallizes out of an aqueous solution, the crystal growth is influenced by the presence of other substances in the solution, such as blood or plant extracts. If a mixture of copper chloride solution with a small amount of whole blood is allowed to crystallize under controlled experimental conditions, an aggregate of crystals forms. Crystallization method can be used as a diagnostic aid to provide information about the systemic conditions and general health of the patient. AIM: This study aims to study the patterns of crystallization and to further determine the efficacy of crystallization test as a screening modality in premalignant lesions and oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: Fifty patients of OSCC, 50 patients of premalignant lesions, and 50 healthy individuals were selected. One drop of blood was collected from the study groups to perform crystallization using cupric chloride. STATISTICAL ANALYSIS USED: Statistical analysis was performed using Chi-square test, Student's t-test (two-tailed), and analysis of variance. RESULTS: The different patterns of crystals formed were studied and statistically analyzed. CONCLUSION: Based on the study, it was concluded that Crystallization test can be used as an effective screening modality for detection of premalignant lesions and OSCC.

Elevated peripheral inflammatory markers are related with the recurrence and canceration of vocal fold leukoplakia.

OBJECTIVE: To evaluate the predictive value of preoperative peripheral inflammatory markers in patients with vocal fold leukoplakia. METHODS: A retrospective study was performed of the patients diagnosed with vocal fold leukoplakia and who accepted carbon dioxide (CO2) laser resection in our center in the last 10 years. We calculated the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and monocyte-to-lymphocyte ratio (MLR) after collecting and analyzing the clinical, histopathological and laboratory data. The potential relation between blood indexes and clinical events as recurrence or canceration was evaluated. RESULTS: A total of 589 patients were involved, including 300 cases without recurrence (group A), 198 with recurrence but not canceration (group B) and 91 transformed into squamous cancer (group C). Baseline analysis of NLR, PLR, and MLR showed no difference among the three groups before the first surgery. But all the indexes significantly elevated in groups B (P < 0.001, < 0.001, 0.023, respectively) and C (P = 0.009, 0.004, 0.007, respectively) in the last operation. The receiver-operating curve (ROC) analysis showed NLR as a potential marker of canceration of leukoplakia (AUC = 0.837) and the cutoff value was 2.505. When regrouping with pathological outcomes, severe dysplasia and squamous cell carcinoma (SCC) groups both revealed a higher level of NLR, PLR, and MLR comparing to the no dysplasia, mild dysplasia, and moderate dysplasia groups. NLR, PLR, and MLR in high-risk group (moderate, severe dysplasia and carcinoma) also elevated comparing to low-risk group (no dysplasia, mild dysplasia) (P = 0.039, 0.011, 0.007, respectively). CONCLUSIONS: The peripheral inflammatory markers NLR, PLR, and MLR are closely connected with the development of vocal fold leukoplakia. NLR may be a potential marker to predict the poor outcomes (recurrence or canceration) of patients in first surgery.

Correlation of serum lactate dehydrogenase and alkaline phosphatase in different histological grades of head and neck squamous cell carcinoma and premalignant lesions.

CONTEXT: Head neck cancer (HNCA) in North-East India accounts for 54.48% cancers at all sites, one of the highest in the country. Increased serum lactate dehydrogenase (LDH) activity is considered as a marker of cellular necrosis while serum alkaline phosphatase (ALP) is recognized as an important marker of induction of tumor cell differentiation. Considering the importance and need of biomarker in HNCA and there being no previous study on tumor markers from this highly prevalent region, we intended to examine the role of serum LDH and ALP in head and neck squamous cell carcinoma (HNSCC) and epithelial precursor lesions (EPLs) and also to find their correlation with the different histological grades of tumor. MATERIALS AND METHODS: This is a prospective observational study on patients with HNSCC and precancerous lesions attending Department of ENT of a Teaching Hospital. Serum LDH and ALP was measured in HNSCC, EPL and control groups and the results were statistically analyzed and compared using analysis of variance, Pearson correlation coefficient, and sensitivity-specificity analysis. RESULTS: Serum LDH was found to be a significant marker of HNSCC. Total serum LDH level was high in both premalignant and HNSCC cases. There was a significant correlation between serum LDH and grades of HNSCC showing highest levels of expression in moderately differentiated SCC. Elevated serum LDH was also seen in erythroleukoplakia, leukoplakia, and verrucous lesion, but serum ALP levels were not significant. CONCLUSIONS: This is the first study from this highly prevalent region of HNCA showing that serum LDH could be regarded as a biomarker for malignant and premalignant conditions of the head and neck.

Comparison of Serum Fucose Levels in Leukoplakia and Oral Cancer Patients.

BACKGROUND: Tumor markers, designated as a broad group of substances produced by malignancies, could be in the form of biochemical substances, immunological substances, cell surface changes and genetic alterations. Cancer, a disorder of cellular behavior is characterized by alteration of serum glycoproteins. L-fucose, a hexose, which is the terminal sugar in most of the plasma glycoproteins, may be useful as a tumor marker for the detection, monitoring and prognostic assessment of malignancies. The aim of the study was to ascertain the role of serum fucose as a biomarker for early detection of oral cancer and to compare serum fucose levels in healthy controls, leukoplakia and oral cancer patients. MATERIALS AND METHODS: The study included 60 (100.0%) subjects, who were grouped as 20 (33.3%) control subjects, 20 (33.3%) squamous cell carcinoma patients and 20 (33.3%) leukoplakia patients. Fucose estimation was done using UV-visible spectrophotometry based on the method as adopted by Winzler using cysteine reagent. The results were analyzed statistically using ANOVA with Bonferroni post hoc tests. RESULTS: Results showed a high significance in serum fucose in oral squamous cell carcinoma (OSCC) and leukoplakia subjects compared to normal controls. There was a gradual increase in the values noted from control to leukoplakia and to squamous cell carcinoma. CONCLUSIONS: Estimation of serum fucose may be a reliable marker and can be used as an effective diagnostic biomarker in oral squamous cell carcinoma patients.

¹H NMR-derived serum metabolomics of leukoplakia and squamous cell carcinoma.

BACKGROUND: Oral cancer (OC) is the sixth commonest cancer worldwide with alarming mortality. If identified at an early stage, the survival rate would be improved. METHODS: We appraised the feasibility of using (1)H nuclear magnetic resonance ((1)H NMR) based metabolomics in the identification of signature metabolites in serum from patients suffering with oral leukoplakia (OLK, n=100), oral squamous cell carcinoma (OSCC, n=100), and healthy control (HC, n=75). (1)H NMR derived data were processed by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) to reveal discriminating metabolites among these groups. Receiver operating characteristic (ROC) curve evaluation was also executed. RESULTS: NMR-derived serum metabolomics reveals eight differentially expressed biomarkers. Among them four biomarkers (glutamine, propionate, acetone, and choline) were able to accurately (ROC; 0.97) segregate 93.5% of OC cases equated to HC with substantial sensitivity and specificity. Similarly, four biomarkers (glutamine, acetone, acetate, and choline) were able to precisely (ROC; 0.96) discriminate, 92.4% of OLK cases from OSCC with considerable sensitivity and specificity. (1)H NMR-based metabolic fingerprint obtained for oral cancer is remarkable, even for OLK stage. CONCLUSION: There is a systemic metabolic response to initial stage of cancer, which carries immense possibility for early appraisal.

Association and prognostic value of serum inflammation markers in patients with leukoplakia and oral cavity cancer.

BACKGROUND: Oral cavity cancer ranks as the fourth leading cancer in men in Taiwan. The development of a serum biomarker panel for early detection and disease monitoring is, therefore, warranted. METHODS: Nine inflammation-associated markers were investigated in 46 patients with leukoplakia, 151 patients with untreated oral cavity squamous cell carcinoma (OSCC), and 111 age- and gender-matched healthy controls using enzyme-linked immunosorbent assay. During a subsequent 28-month surveillance of OSCC patients, serum samples were prospectively collected at predetermined intervals following the completion of therapy. RESULTS: Logistic regression analysis showed matrix metalloproteases (MMP)-2, MMP-9, C-reactive protein (CRP), transforming growth factor-ß1 (TGF-ß1), and E-selectin having the best discrimination power between groups and significant elevation trends of those five markers were noted from control to OSCC. By combining those five markers, a 0.888 and 0.938 area under curve by ROC curve analysis with 67.4% and 80% overall sensitivity and fixed 90% specificity for leukoplakia and OSCC groups were demonstrated. In the follow-up period, 25 OSCC patients developed recurring or secondary tumors. All examined markers had decreased in relapse-free patients following treatment. However, in patients with relapse, interleukin-6, CRP, and serum amyloid A remained at elevated levels. Statistical analysis showed that patients with CRP ≧2 mg/L and E-selectin ≧85 ng/mL at baseline had highest probability of relapse (odds ratio=3.029, p<0.05). CONCLUSIONS: The results indicate that inflammation plays a crucial role in the pathogenesis process of OSCC. By examining the inflammation markers, physicians could potentially identify patients at risk of cancer transformation or relapse.

Serum lipid profile in oral cancer and leukoplakia: correlation with tobacco abuse and histological grading.

INTRODUCTION: Role of alterations in serum lipid profile in oral cancer remains controversial. The present study aimed to evaluate the implications of altered serum lipid profile in patients with oral cancer (OC), oral leukoplakia (OLP), and tobacco habits. MATERIALS AND METHODS: Thirty patients with OC, 30 with OLP, 30 tobacco abusers (TAs), and 30 age and sex matched healthy controls were included in the study. Serum lipid profile including total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), Very low density lipoprotein (VLDL), and triglycerides (Tg) were evaluated using a fully automatic Biochemistry analyzer. Difference in lipid profile in various types of TA, that is, smokeless tobacco (SLT), smoking tobacco (ST), and a combination (Comb) usage of both forms were also analyzed. RESULTS: TC, HDL, and LDL were much lower in the OC group compared with control. Although these parameters were low in the OPC group compared with controls, the difference was not significant. On histological analysis, TC and HDL were found to decrease marginally with loss of tumor differentiation in OC. No correlation was found between the mean serum lipid profiles and degree of dysplasia in OLP. TC and HDL were significantly lesser in all forms of TA when compared with control. CONCLUSIONS: There may be an inverse relationship between serum lipid profile and OC. No significant reduction in lipid profile was observed in the OLP group. This may indicate that hypolipidemia is a late change occurring during carcinogenesis or is an effect rather than the cause of cancer.

Correlation of trace elemental profiles in blood samples of Indian patients with leukoplakia and oral submucous fibrosis.

In order to ascertain possible correlation between alterations in trace elemental profile and the progression or regression of two most common potentially malignant disorders affecting oral cavity, namely oral submucous fibrosis and leukoplakia, blood from 60 patients from each group of patients as well from 30 healthy individuals was analyzed for elemental profiling employing EDXRF technique. Out of the 16 detected elements (K, Si, Ca, V, Cr, Ni, Mn, Fe, Cu, Zn, Se, Br, Rb, Sr, Co, and Pb), Mn, Fe, Zn, Br, and Co showed remarkable alteration in their profile in both leukoplakia and oral submucous fibrosis patients with respect to the normal healthy individuals. While Zn, Br, and Fe reflected similar changes--showing gross depletion in both the diseased groups, Mn and Co depicted inverse pattern of alterations in their concentrations in the two types of precancerous disorders when compared to the control subjects.

[Leucoplakia vesicae in females: diagnosis and treatment].

Sixty patients suffering from leucoplakia vesicae (LV) were examined using cystoscopy with biopsy of the urinary bladder wall, blood enzyme immunoassay for detection of antibodies to agents of sexually transmitted infections (STI), uroflowmetry, culture analysis of cervical canal and mucosa samples for STI. As shown by a pathomorphological examination of the vesical mucosa biopsy specimens, long-term persistence of pathogenic (chlamydia, trichomonades) and opportunistic (mycoplasma, ureaplasma, fungi) flora underlies development of LV. Morphogenesis of LV is characterized by hyperplastic reactions of urothelium and its metaplasy in laminated squamous keratosic epithelium, often with para- and dyskeratosis, developing in the presence of inflammatory reactions in the lamina in the presence of persisting infection. In LV, specific infection agents are often found in the urogenital tract. The spectrum of these agents is identical for samples from the cervical canal and vesical mucosa from leucoplakia foci. Vesical mucosa is most frequently contaminated with Mycoplasma hominis (57.2%), Candida albicans (51.4%), Ureaplasma urealiticum (37.1%) and Trichomonas vaginalis (22.9%). Associations of the infection agents are detected in 70% of LV patients. Persistent dysuria is a basic clinical symptom of leucoplakia. The following therapeutic measures should be taken: transurethral coagulation of the vesical mucosa, intravesical therapy, immunocorrection, antibacterial treatment by standard schemes or according to the isolated flora sensitivity.

Serum levels of folate, homocysteine, and vitamin B12 in head and neck squamous cell carcinoma and in laryngeal leukoplakia.

BACKGROUND: The authors evaluated serum levels of folate, homocysteine, and vitamin B(12) in patients with head and neck squamous cell carcinoma (HNSCC) and in patients with laryngeal leukoplakia, a well known preneoplastic lesion. METHODS: One hundred forty-four consecutive, untreated patients with HNSCC and 40 consecutive, untreated patients with laryngeal leukoplakia were enrolled in the Department of Otolaryngology at the authors' institution. Data from those patients were compared with data from one control group, which included 90 smokers, and from another control group, which included 120 nonsmokers. Serum levels of homocysteine, folate, and vitamin B(12) were measured by an automated immunoassay method based on fluorescence polarization immunoassay technology. RESULTS: Comparing groups by Student-Newman-Keuls test, serum folate levels were significantly lower in patients with HNSCC and in patients with laryngeal leukoplakia compared with serum folate levels in both the smoker control group and the nonsmoker control group. Serum homocysteine levels in patients with HNSCC were significantly higher compared with homocysteine levels both in the smoker and nonsmoker control groups and in patients with laryngeal leukoplakia. There were no statistically significant differences between groups in serum vitamin B(12) levels. CONCLUSIONS: A role for folate deficiency as a risk factor in head and neck carcinogenesis is plausible. A chemoprevention protocol with folate is both feasible and ethically correct and is in progress at the authors' institution. Homocysteine levels in patients with HNSCC probably are affected largely by the HNSCC phenotype. An accumulation of homocysteine may reveal a genetic defect, which, theoretically, may be a target for pharmacologic therapy, for example, with antifolic drugs.

[Disorders of hormonal homeostasis in patients with kraurosis and leukoplakia vulvae]. / Narusheniia gormonal'nogo gomeostaza u bol'nykh kraurozom i leikoplakiei vul'vvy.

A study was made into the functional status of hypophysis, adrenal cortex, ovaries, and thyroid gland in patients with kraurosis and leukoplakia vulvae. The above patients demonstrated profound disturbances in the system hypophysis-ovaries, that attest to the need for applying corrective therapies to treat the disturbed hormonal homeostasis.

[Ferritin (tumor marker) in patients with laryngeal cancer and precancerous conditions]. / Ferrytyna (marker nowotworowy) u chorych na raka krtani w stanach przedrakowych.

Evaluation of the ferritin as a neoplasm marker was performed in laryngeal cancer patients. In 35 laryngeal cancer patients and in 21 laryngeal leucoplakia patients the ferritin serum level and the blastic transformation of leucocytes were tested. In both groups the high ferritin level was discovered. It is the sign of low specificity of ferritin as a neoplasm marker. There was no correlation between the ferritin serum level and the degree of the blastic changes of leucocytes.

[Endogenous factors in the development of leukoplakia and kraurosis vulvae]. / Nekotorye éndogennye faktory v razvitii leikoplakii i krauroza vul'vy.

Levels of tropic hormones of the pituitary, glucocorticoids and male and female sex hormones were assayed in 17 cases of kraurosis and leukoplakia of the vulva prior to treatment. Most patients revealed an abnormally high androgen/estrogen ratio, elevated level of glucocorticoids and low concentrations of thyrotropic hormone, prolactin and T3. The above disturbances are closely related and interdependent.

Serum retinol level in patients with pre and malignant lesions of female genital tract.

The blood retinol level in women with genital tract malignancies and in a control group has been determined. In all groups of patients the mean serum retinol level was significantly lower than in control group. Based on these results we can postulate, that the low retinol level could predispose to the development of malignancy in female genital tract.

Detection of A, B and H(O) antigens in normal and neoplastic epithelium of the urinary bladder by the specific red cell adherence test (SRCA).

A, B and H(O) blood group antigens in some tissues are readily demonstrable by the specific red cell adherence test (SRCA) which is a modification of Coomb's mixed cell adherence technique. Paraffin-embedded tissue sections obtained from tumors of the urinary bladder were submitted to SRCA. Normal bladder epithelium adjacent to neoplastic lesions and the leukoplakia were found to contain the three antigens. The antigens were completely absent in 18 cases and partially present in one of the bladder carcinoma. It was also suggested that the prognosis of the tumor correlates with the antigen loss. SRCA is sensitive, reproducible and technically easy for detection of tissue antigens, and it would be expected to play a crucial role for demonstration of immunological dedifferentiation in tissues which normally keep the antigens.